Journal: bioRxiv

Article Title: Glycopolymer-conjugated antigens as an inverse vaccine platform prevent anaphylaxis in a pre-clinical model of food allergy

doi: 10.1101/2023.03.23.534004

Figure Lengend Snippet: Experimental schema. B. Change in core body temperature at indicated time points following challenge with BLG in BLG + CT sensitized mice. The mice were given antibiotic water throughout the experiments, and were treated with either PBS ( n = 8), unmodified BLG ( n = 10), or BLG-p(Man) ( n = 8) with two doses before sensitization. C. mMCP-1 from post-challenge serum of mice in B. D-F. BLG-specific IgG (D), IgG1 (E), and IgE (F) from in mice serum collected three days before the oral challenge. G . Fold-increase of GATA3 + or IL-4 + CD4 + T cells of splenocytes from mice sacrificed two weeks post-challenge, and restimulated with free BLG for 6 hr. H. LegendPlex analysis from culture supernatants of splenocytes from mice sacrificed two weeks post-challenge and stimulated for 4 days with BLG. Statistical differences determined by one-way (C-F) or two-way (G, H) ANOVA using Tukey’s post hoc test. Data were pooled from two experiments. Data represent mean ± SEM, *p < 0.05, **p<0.01.

Article Snippet: For the BLG-specific IgE ELISA, serum from the sensitized mice was diluted and added to BLG-coated 96-well ELISA plates (Costar highbind flat-bottom plates, Corning).

Techniques:

Journal: bioRxiv

Article Title: Glycopolymer-conjugated antigens as an inverse vaccine platform prevent anaphylaxis in a pre-clinical model of food allergy

doi: 10.1101/2023.03.23.534004

Figure Lengend Snippet: Experimental design. C3H/HeJ mice were given antibiotic water throughout the experiments and were s.c. administered with two doses of either saline, unmodified BLG, or BLG-pMan before sensitization. The mice were then orally challenged with BLG to assess the allergic reactions. B, Change in core body temperature at indicated time points following challenge with BLG in BLG + CT sensitized mice. C. BLG-specific IgG, IgG1, and IgE in mice serum collected three days before the oral challenge. D. mMCPT-1, IL-6, and IL-4 from post-challenge serum of mice in B. E. Th2 cytokine (IL-5 and IL-13) levels from culture supernatants of splenocytes from mice sacrificed one week post-challenge and stimulated for 4 days with BLG. Data were pooled from three experiments. Statistical differences determined by one-way ANOVA using Dunnett’s post hoc test (B-D) or Kruskal-Wallis test (E). (*p < 0.05, **p<0.01).

Article Snippet: For the BLG-specific IgE ELISA, serum from the sensitized mice was diluted and added to BLG-coated 96-well ELISA plates (Costar highbind flat-bottom plates, Corning).

Techniques:

Journal: bioRxiv

Article Title: Glycopolymer-conjugated antigens as an inverse vaccine platform prevent anaphylaxis in a pre-clinical model of food allergy

doi: 10.1101/2023.03.23.534004

Figure Lengend Snippet: Experimental design. C3H/HeJ mice were given antibiotic water and sensitized weekly by intragastric gavage of 0.1 mg/g body weight of BLG plus 6.7 mg/g body weight of the mucosal adjuvant cholera toxin. 5 days after sensitization, the mice were i.p. injected with anti-CD20 antibody or isotype control, and two days later followed by s.c. administration of two doses of saline, free BLG, or BLG-p(Man). The mice were then challenged i.g. with BLG to assess their anaphylactic responses (except for the free BLG group, in which more than half of mice died from anaphylactic reactions upon treatment). Mice treated with anti-CD20 and saline, or BLG-p(Man) were given weekly low doses of BLG (1 mg per mouse) and challenged again afterwards. Allergic mice were defined as core body temperature dropping by 1°C or more upon BLG challenge. B . The core body temperature at 30 min after each therapeutic injection. C. Change in core body temperature at indicated time points following oral challenge with BLG. D. mMCPT-1 measured from post-challenge serum of mice in C. E, F. BLG-specific IgG (E) and IgE (F) in the mouse serum before and after treatment. G. Change of the ratio of BLG-specific IgG to IgE before and after the treatment. H. Change in core body temperature of mice kept in weekly dosing of BLG after treatment, following a second oral challenge with BLG. Data were pooled from two experiments. Statistical differences determined by one-way ANOVA using Dunnett’s post hoc test (B), Kruskal-Wallis test (D), two-way ANOVA using Bonferroni’s post hoc test (E, F), or Wilcoxon test (G) (*p < 0.05, **p<0.01, ***p<0.001, ****p<0.0001).

Article Snippet: For the BLG-specific IgE ELISA, serum from the sensitized mice was diluted and added to BLG-coated 96-well ELISA plates (Costar highbind flat-bottom plates, Corning).

Techniques: Injection